Thesis (D.Phil.)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: Chapter 1 provides literature based background information on the clinical
importance of sperm morphology as recorded by strict criteria during the
diagnostic approach of the infertile couple. Furthermore, the use of a sequential
diagnostic schedule for couples in an assisted reproductive programme is
emphasized. The author revisited the literature on chromatin packaging of
spermatozoa and addresses this issue as an additional semen parameter
providing information relating to DNA damaged spermatozoa. The chapter also
includes evidence underlining the growing need for the implementation of the
acrosome reaction as an important contribution to the assisted reproductive
programme. Chapter 2 provides detailed descriptions of the material and
methods used during the study. Chapter 3 is sub-divided into 5 sections, each of
which represents a separate study that was prepared as a scientific paper. The
study included 338 couples consulting for infertility treatment at various
gynaecologists in Pretoria and Johannesburg. The diagnostic assisted
reproductive laboratory support was provided by the Andrology laboratory of Drs
du Buisson and partners from Pretoria. In the first study the role of chromatin
packaging as an indicator of in vitro fertilization rates, the semen samples from
72 men were used to record their chromatin packaging quality as well as their
sperm morphology classification. Significant different percentages CMA3staining
(mean±SE) were recorded among the 2 morphology groups, namely 65.9%±3.5
and 44.5%±1.7 (p=0.001). Using cut off values for chromatin packaging
established during the first study, the second study utilized semen from 140 men in the in vitro fertilization (IVF) and intracytoplasmic sperm injection programme
(ICSI) to analyze for sperm concentration, motility, morphology and chromatin
packaging (CMA3).IVF and ICSI data were stratified using 3 basic cut off values
for CMA3staining, namely <44%, >44-60% and >60%. The study concluded that
results on the chromatin packaging quality of spermatozoa could be used as an
additional parameter of sperm quality since it could provide valuable information
on decondensation status of a given sperm population. The third study aimed to
establish zona pellucida induced acrosome reaction response (ZIAR) among 35
couples with normal and G-pattern sperm morphology and repeated poor
fertilization results during assisted reproduction treatment. Interactive dot
diagrams, divided patients into 2 groups i.e. ZIAR<15% and ZIAR>15% with
mean fertilization rates of 49% and 79%, respectively. The study concluded that
the ZIAR test has diagnostic potential, since it can assist the clinician to identify
couples that will benefit from ICSI therapy. The forth study revisited the
importance of micro-assay for acrosome reaction determinations in a diagnostic
andrology laboratory. The micro-assay not only allows the use of a single zona
pellucida, but also facilitates the future possibility of using recombinant zona
pellucida proteins in a diagnostic test system. The final study in Chapter 3
includes results obtained from 49 couples (172 oocytes) and aimed to evaluate
the role of chromatin packaging and sperm morphology during sperm-zona
binding, sperm decondensation and the presence of polar bodies among 170
oocytes that failed in vitro fertilization (IVF). Odds ratio analyses indicated that
being in the a group with elevated CMA3 staining i.e. >60%, the risk of decondensation failure increases 15.6 fold relative to normal CMA3 staining
<44%. Chapter 4 underlines the validity of the sequential diagnostic approach
and summarizes the results and value of a multistep diagnostic scheme. The
chapter concludes with the recommendation that both chromatin packaging
quality and zona pellucida mediation of the acrosome reaction should be part of
the diagnostic tools in the assisted reproductive programme. / AFRIKAANSE OPSOMMING: Die literatuuroorsig in Hoofstuk 1 konsentreer in hoofsaak op die kliniese belang
van sperm morfologie en die uitbreiding van die diagnostiese toetse en hantering
van die egpaar in die reproduktiewe ondersteuningsprogram. Die kromatien
pakkingskwaliteit van die spermsel word onderskryf as In belangrike toevoeging
tot die diganostiese program, aangesien ONS skade dikwels saam met
kromatiendefekte aangetref word. Die rol van die akrosoomreaksie word ook in
detail literatuuroorsigtelik beklemtoon. Hoofstuk 2 bevat volledige inligting
omtrent materiaal en metodes wat in die studie gebruik is. Hoofstuk 3 bevat die
eksperimentele gegewens wat in 5 afsonderlike sub-afdelings as wetenskaplike
publikasies aangebied word. Die studies bestaan uit data van 338 pasiënte, wat
deur verskillende ginekoloë van Pretoria en Johannesburg gekonsulteer is
waartydens drs. du Boisson en vennote van Pretoria die diagnostiese
reproduktiewe laboratoriumdienste verskaf het. Die eerste studie stel dit ten doel
om die belang en korrelasie van die spermsel kromatienpakkingskwaliteit van 72
mans te vergelyk met die morfologiese bou van sie sel. Aangesien morfologie
reeds gevertig is as 'n kliniese voorspeller van bevrugting was dit nodig om
hierdie parameter te vergelyk met die kromatienpakking van die sel. Twee
afsnypunte word vir die normo-en teratozoospermiese mans identifiseer naamlik,
44.5%±1.7 en 65.9%±3.5 (p=O.001),respektiewelik. Die tweede studie gebruik
die afsnypunte 44% en 66% om die in vitro bevrugting en intrasellulêre sperm
inspuiting (ICSI) data te ontleed. Die resultate dui aan dat kromatienpakking In
waardevolle bydrae tot die diagnostiek van die pasiënte lewer. Die derde studie stel dit ten doelom die waarde van die zona pellucida geinduseerde
akrosoomreaksie (ZIAR) te bepaal. Die studie sluit die data van 35 egpare in wat
almal normale of G-patroon morfologie het en verder onverklaarde swak
bevrugtings resultate tydens in vitro bevrugtingsterapie. Interaktiewe punt
diagram (interactive dot diagrams) verdeel die data in twee groepe naamlik,
ZIAR<15% en ZIAR>15% met gemiddelde bevrugtingssyfers van 49% en 79%,
respektiewelik. Die studie sluit af met die gedagte dat die ZIAR toets 'n groep
pasiënte identifiseer met 'n besondere fisiologiese afwyking d.i. subnormale
akrosoom respons op zona pellucida blootstelling. Die vierde afdeling van die
hoofstuk onderstreep die belang van die mikro-tegniek vir die bepaling van die
akrosoom reaksie, wat tydens die projek gebruik is Die vyfde afdeling van
Hoofstuk 3 stel dit ten doelom 170 onbevrugde eierselle van 49 pasiënte te
ontleed vir moontlike oorsake vir die mislukte bevrugting. Ondersoeke sluit in die
kromatienpakking, sperm-zona binding, sperm dekondensasie en die
teenwoordigheid van polêre liggaampies. Statisties blyk dit dat indien 'n
kromtienpakking nie normaal is nie (>66%) het die spermsel 'n 15 keer groter
kans om nie te dekondenseer nie. Hoofstuk 4 bespreek die noodsaaklikheid van
die diagnostiese skedule by die hantering van die onvrugbare egpaar in.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/51752 |
| Date | 12 1900 |
| Creators | Esterhuizen, Aletta Dorothea |
| Contributors | Franken, D. R., Stellenbosch University. Faculty of Medicine & Health Sciences. Dept. of Medicine. |
| Publisher | Stellenbosch : Stellenbosch University |
| Source Sets | South African National ETD Portal |
| Language | en_ZA |
| Detected Language | English |
| Type | Thesis |
| Format | 159 p. |
| Rights | Stellenbosch University |
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